日本細胞生物学会
HOME- Home
- >
- Experimental protocol top
- >
- Immunofluorescence by Zenon
Immunofluorescence by Zenon
＊download PDF| Author | Akihiro Harada | ||
|---|---|---|---|
| Affiliation | Department of Cell Biology, Graduate School of Medicine, Osaka University | ||
| Contact | inquiry about this protocol | ||
| Home Page | http://www.med.osaka-u.ac.jp/pub/acb/eng.html | ||
| Keyword | |||
| Published | 2015-08-26 | Last Update | 2015-08-26 |
| View | 25 | download PDF | 7 |
Summary
Methods
Detail ＊Click photo to see a large size.
- Wash the sections with PBS once
- Block the nonspecific staining by treating the sections in 5％Normal donkey Serum/PBS
- Treat the sections with diluted primary antibody in 5％Normal donkey Serum/PBS, 37C 1hr
- Wash the sections with PBS several times
- Block the nonspecific staining by treating the sections in 5％Normal donkey Serum/PBS
- Treat the sections with diluted secondary antibodies in 5％Normal donkey Serum/PBS, 37C 30min
- Wash the sections with PBS several times
- Block the nonspecific staining by treating the sections in 5％Normal Goat Serum/PBT(0.1％TritonX-100/PBS)
- Treat the sections with diluted tertiary antibodies (labeled by Zenon) in PBT, RT 1hr
- Wash the sections with PBS several times
- Postfix the sections with 4％PFA/PBS,RT 15min
- Wash the sections with PBS once
- Embedding
- Protocol for making tertiary antibody
1) Mix antibody solution 10μℓ+Zenon Comp A 10μℓ, RT, more than 5min（less than a few days）
2) Add Zenon Comp B 10μℓ to mixture 1（total 30μℓ）, RT 5min
3) Add PBT 70μℓ to mixture 2 (total 100μℓ)
